9alpha halo delta4 androstenes



90: HALO A ANDROSTENES Gordon H. Thomas, New Brunswick, and Richard W.

Thoma, Somerville, N. J., assignors to Olin Mathieson ChemicalCorporation, New York, N. Y., a corporation of Virginia I No Drawing.Application October 29, 1957 7 Serial No. 693,039

7 Claims. (31. 260-39145 This invention relates to the synthesis ofvaluable steroids and, more particularly, to the preparation of newsteroids of the general formula wherein R is hydrogen, R is S-hydroxy ortogether R and R is keto, X is halogen (preferably fluoro), and Y ishydrogen or acyl, particularly the acyl radical of a hydrocarboncarboxylic acid of less than ten carbon atoms.

These new steroids are physiologically active substances which possessandrogenic activity. Thus, they may be administered instead of and inthe same manner as testosterone propionate, for example, in thetreatment of hypogonadism, for which purpose they are formulated inparenterally acceptable preparations (e. g., aqueous suspension).

The starting materials for the steroids 01? this invention include:9a-halo-l1B-hydroxyandrostenediones (e. g., 9a-fiuoro-A-androsten-1lfl-ol-3,17-dione and 9a-chloro- A-androsten-11/8-ol-3,17-dione), and 9a-halo-11-ketoandrostenediones (e.g., 9a-fluoro-A -androstene-3,11,17- trione and 9a-chloro-A-androstene-3,11,17-trione), These starting materials can be prepared bythe method disclosed in the U. S. application of Josef Fried, Serial No.469,848, filed November 18, 1954. To prepare the steroids of thisinvention, wherein Y is hydrogen, such starting materials are subjectedto the action of the enzymes of Streptomyces roseochromogenus WaksmanNo. 3689 (Institute of Microbiology. Rutgers University, New Brunswick,New Jersey) in an aqueous-medium containing a source of nitrogenousfactors and an assimilable source of carbon and energy, in the presenceof oxygen, and the 16a-hydroxy steroid formed is recovered.

The resulting 16a-hydroxy steroid (e. g., 9a-fluoro-A androstene115,160: diol-3,l7-dione and 9a-fluoro-M-androsten-16e-ol-3,11,17-trione) can then be esterified, if desired, inthe usual manner, as by treatment with the acid anhydride-or acyl halideof the desired acid in the presence of an organic base, such aspyridine. Among the suitable acid anhydrides and acyl halides may bementioned the acid anhydride and acyl chlorides of hydrocarboncarboxylic acids of less than ten carbon atoms, as exemplified by thelower alkanoic acids (e. gpacetic, propionic and caproic acid),monocyclic aromatic carboxylic acids (e. g., benzoic and m-toluic acid),monocyclic aralkanoic acids (e. g., phenacetic and B-phenylpropionicacid), cycloalkanecarboxylic acids (e. g., cyclohexanecarboxylic acid),lower alkenoic acids, and lower cycloalkenoic acids.

2,853,502 Patented Sept. 23, I958 "ice The following examples areillustrative of the invention (all temperatures being in centigrade):

EXAMPLE 1 9a-flu0r0-] 1B,16a-dihydroocyandrostenedionie (a)Fermerrmtion.Eight flasks, each containing 5 0 ml. of the followingsterilized medium (A):

Distilled water to make one liter.

are inoculated each with 0.8 ml.of a suspension obtained by adding 8 ml.of 0.01% aqueous Duponal (a wetting agent) solution to a three-week-oldslant culture of Streptomyces roseachromiogenus'WaksmanNo. 3689 onBennetts agar (yeast extract, 1%; beef extract, 1%; NZ Amine A, 2%;glucose, 10%; agar, 15%). After 66 hours of incubation at 25 with rotarymechanical agitation (280 R. P. M., 2 inch radius), a 10% (vol/vol.)transfer is made to 48 flasks containing 50 ml. portions of the samesterile medium A. A total of 720 mg. of9afluoro-l1fi-hydroxyandrostenedione is added aseptically in methanolsolution containing 15 mg. of steroid/ml. The

flasks are incubated for 96 hours and the contents of they flaskspooled. The contents are then adjusted to pH 4.1 with 12 N sulfuric acidand filtered through a Seitz clarifying pad with suction. The flasks andpad are washed with about five 50 ml. portions of water. The combinedvolume of filtrate and washings is about 2630 ml. The combined filtrateand washings is held at 15 with 100 ml. of chloroform. Paperchromatographic analyses indicates that no starting material remains.

(b) Isolation-The culture filtrate and wash is extracted four times withone liter of chloroform, the combined chloroform extracts are Washedwith water, dried over sodium sulfate and evaporated to dryness invacuo. Trituration of the residue with hexane yields about 440 mg. of9a-fluoro-11,8,l6a-dihydroxyandrostenedione having a M. P. of about249-268. Crystallization 1 from chloroform-methanol gives a pure samplemelting at about 280-282; [0&1 +173 (c., 0.97 in chloroform);

M,};-, 237 m (6 16,200) A213? 2.86, 2.95, 3.10, 5.72 6.05,u.

AnaZysis.-Calcd. for C H O F: C, 67.83; H, 7.49; F, 5.65.. Found: C,67.70; H, 7.49; F, 5.74.

In a similar manner, by substituting 9oc-ChlO1'O-1lfl-hY-droxyandrostenedione for the 9a-fluoro-11 3-hydroxyandrostenedione inthe procedure of Example 1, 9a-chloro- 1 13,16ct-dihydroxyandrostenedione is formed.

EXAMPLE 2 9a-fluoro-M-androstene-l 6a-0l-3,11,1 7-tri0ne Following theprocedure of Example 1, but substituting 720 mg. of 9a-fluoro-A-androstene-3,11,17-trione for the 9a-fluoro-11fi-hydroxyandrostenedione, 9a-fluoro-A -anClI0StI1C-l6oc-Ol-3,1l,17-lfl01'16 is obtained.

Similarly, upon substitution of 9a-chloro-A -androstene- 3,11,17-trionefor the 9a-fluoro steroid in Example 2,9achloro-M-androstene-l6a-ol-3,11,17-trione is formed.

EXAMPLE 3 To a solution of 50 mg. of9a-fluoro-11/3,16o-dihydroxyandrostenedione in 1 ml. of pyridine isadded 0.3 ml. of propionic anhydride, and the resulting solution isallowed to remain at room temperature for 18 hours. The mixture is thendiluted with water, the precipitated solid col- 3 lected, washed withWater, dried andcrystallized from acetone-hexane.

Similarly, if 9u-chloro-11,8,16a-dihydroxyandrostenedione, 9a-fiuoro-A-androstene-16a-ol-3,11,17-trione, and 9a-chloro-A-androstene-l6a-ol-3,11,17-trione are substituted for the9a-fluoro-115,16u-dihydroxyandrostenedione in the procedure of Example3, the respective l6a-propionate esters are formed.

Furthermore, if another acylating agent such as acetic anhydride orbenzoyl chloride is substituted for the propionic anhydride in theprocedure of Example 3, the respective 16a-este1' is obtained.

The invention may be otherwise variously embodied within the scope ofthe appended claims.

What is claimed is:

,1. A steroid of the general formula '4, wherein R is hydrogen, R isfi-hydroxy and together R and R is keto, X is halogen, and Y is selectedfrom the group consisting of hydrogen and the acyl radical of ahydrocarbon carboxylic acid of less than ten carbon atoms.

2. 9a-halo-A -androstene-1 1/3,16a-diol-3,17-dione.

3. 9a-fluoro-A -androstene-1 1B,16oc-diOl-3,17di0118.

4. The 1600 ester of 9m-halo-A -androstene-115,16mdiol-3,-l7-dione witha hydrocarbon carboxylic acid of less than ten carbon atoms.

5 9oz fiuoro A androstene llfl,l6a diol 3,17 dione l6u-propionate.

6. 9a-halo- A -androstene-16u-ol-3,1 1,17-trione.

7. 9a-fluoro-A -androstene-16a-ol-3,11,17-trione.

References Cited in the file of this patent UNITED STATES PATENTS2,773,080 Bernstein Dec. 4, 1956

1. A STEROID OF THE GENERAL FORMULA